Biocompatible implantable sensor apparatus and methods

ABSTRACT

Enzymatic and non-enzymatic detectors and associated membrane apparatus, and methods of use, such as within a fully implantable sensor apparatus. In one embodiment, detector performance is controlled through selective use of membrane configurations and enzyme region shapes, which enable accurate detection of blood glucose level within the solid tissue of the living host for extended periods of time. Isolation between the host&#39;s tissue and the underlying enzymes and reaction byproducts used in the detectors is also advantageously maintained in one embodiment via use of a non-enzyme containing permeable membrane formed of e.g., a biocompatible crosslinked protein-based material. Control of response range and/or rate in some embodiments also permits customization of sensor elements. In one variant, heterogeneous detector elements are used to, e.g., accommodate a wider range of blood glucose concentration within the host. Methods of manufacturing the membranes and detectors, including methods to increase reliability, are also disclosed.

PRIORITY AND RELATED APPLICATIONS

This application is a divisional of and claims priority to co-owned,co-pending U.S. patent application Ser. No. 15/170,571 filed on Jun. 1,2016 of the same title, which is incorporated herein by reference in itsentirety. Additionally, this application is related to co-owned andco-pending U.S. patent application Ser. No. 13/559,475 filed Jul. 26,2012 entitled “Tissue Implantable Sensor With Hermetically SealedHousing,” and co-pending and co-owned U.S. patent application Ser. No.14/982,346 filed Dec. 29, 2015 and entitled “Implantable SensorApparatus and Methods,” each of the foregoing incorporated herein byreference in its entirety. This application is also related to U.S.patent application Ser. No. 10/719,541 filed Nov. 20, 2003, now issuedas U.S. Pat. No. 7,336,984 and entitled “Membrane and ElectrodeStructure for Implantable Sensor,” also incorporated herein by referencein its entirety.

GRANT INFORMATION

This invention was made in part with government support under NIH GrantNo. DK-77254. The United States government has certain rights in thisinvention.

COPYRIGHT

A portion of the disclosure of this patent document contains materialthat is subject to copyright protection. The copyright owner has noobjection to the facsimile reproduction by anyone of the patent documentor the patent disclosure, as it appears in the Patent and TrademarkOffice patent files or records, but otherwise reserves all copyrightrights whatsoever.

1. TECHNICAL FIELD

The disclosure relates generally to the field of sensors, therapydevices, implants and other devices which can be used consistent withhuman beings or other living entities for in vivo detection andmeasurement or delivery of various solutes, and in one exemplary aspectto methods and apparatus enabling the use of such sensors and/orelectronic devices for, e.g. monitoring of one or more physiologicalparameters, including through use of a novel membrane structure.

2. DESCRIPTION OF RELATED TECHNOLOGY

Implantable electronics is a rapidly expanding discipline within themedical arts. Owing in part to great advances in electronics andwireless technology integration, miniaturization, and performance,sensors or other types of electronics or implantable devices (e.g.,therapy agent delivery devices or materials, implants, and the like)which once were beyond the realm of reasonable use in vivo on a livingsubject can now be surgically implanted within such subjects withminimal effect on the recipient subject, and in fact many inherentbenefits.

One particular area of note relates to blood glucose monitoring forsubjects, including those with so-called “type 1” or “type 2” diabetes.As is well known, regulation of blood glucose is impaired in people withdiabetes by: (1) the inability of the pancreas to adequately produce theglucose-regulating hormone insulin; (2) the insensitivity of varioustissues that use insulin to take up glucose; or (3) a combination ofboth of these phenomena. To correct this disregulation requires bloodglucose monitoring.

Currently, glucose monitoring in the diabetic population is basedlargely on collecting blood by “fingersticking” and determining itsglucose concentration by conventional assay. This procedure has severaldisadvantages, including: (1) the discomfort associated withfingersticking, which should be performed repeatedly each day; (2) thenear impossibility of sufficiently frequent sampling (some blood glucoseexcursions require sampling every 20 minutes, or more frequently, toaccurately treat); and (3) the requirement that the user initiate bloodcollection, which precludes warning strategies that rely on automaticearly detection. Using the extant fingersticking procedure, the frequentsampling regimen that would be most medically beneficial cannot berealistically expected of even the most committed patients, andautomatic sampling, which would be especially useful during periods ofsleep, is not available.

Implantable glucose sensors have long been considered as an alternativeto intermittent monitoring of blood glucose levels by the fingerstickmethod of sample collection. These devices may be partially implanted,where certain components reside within the body but are physicallyconnected to additional components external to the body via one or morepercutaneous elements. Partially implanted sensors (discussed in greaterdetail below) are not viable for long-term use, particularly due to theundesirability of having an essentially open wound on the body for anextended period, and all of the attendant problems associated therewith(including greater risk of infection, the body's natural response toattempt to expel the percutaneous or “through the skin” portion of theimplant, etc.).

Implantable sensor devices may alternatively be fully implanted, whereall components of the system reside within the body, and there are nopercutaneous elements. The operability of one such fully implantedsensor has been demonstrated as a central venous implant in dogs (Armouret al., Diabetes, 39:1519 1526 (1990), incorporated herein by referencein its entirety). Although this sensor provided direct recording ofblood glucose, which is most advantageous for clinical applications, thedescribed implantation at a central venous site poses several risks anddrawbacks, including risk of blood clot formation and vascular walldamage. An alternative that does not present such risks to the user isto implant the sensor in e.g., a “solid” tissue site and to relate theresulting signal to blood glucose concentration.

Typical sensors implanted in solid tissue sites measure theconcentration of solutes, such as glucose, in the blood perfusing themicrocirculation in the vicinity of the sensor. Glucose diffuses fromnearby capillaries to the sensor surface. Because such diffusion occurseffectively only over very small distances, the sensor responds to thesubstrate supply only from nearby blood vessels. Conversely, solutesthat are generated in the locality of the sensor may be transported awayfrom the sensor's immediate vicinity by the local microvasculature. Ineither case, the local microcirculation may influence the sensor'sresponse.

Optical glucose sensors are known in the prior art. Schultz and Mansouridisclosed one such version of an optical sensor (J. S. Schultz and S.Mansouri, “Optical Fiber Affinity Sensors,” Methods in Enzymology, K.Mosbach, Ed., Academic Press, New York, 1988, vol. 137, pp. 349-366). Avariety of other optical techniques including optical coherencetomography, near infrared spectroscopy, Raman spectroscopy, andpolarimetry have been tried and failed. Light-based systems using eitherabsorption of light, or emission of light when glucose is “excited” bylight have not proven to be accurate since there is no specific lightabsorption or emission spectrum for glucose. Furthermore, numerous otherchemicals or interfering substances in the blood overlap in spectrumwith glucose, causing optical methods to be insufficiently specific forglucose monitoring.

A number of electrochemical glucose sensors have also been developed,most of which are based on the reaction catalyzed by glucose oxidase.One such configuration involves the use of glucose oxidase to catalyzethe reaction between glucose and oxygen to yield gluconate and hydrogenperoxide. The hydrogen peroxide is either detected directly, or furtherdecomposed by a second enzyme, e.g. catalase, in which case the sensormeasures oxygen consumption. In order for glucose oxidase-based sensorsto function properly, the presence of excess molecular oxygen relativeto molecular glucose is necessary. However, this requirement gives riseto a sensor design problem related to “oxygen deficit,” since theconcentration of oxygen in subcutaneous tissue is significantly lessthan that of glucose.

For example, the typical concentration of glucose in the blood is about4 to about 20 mM, whereas a typical concentration of oxygen in bloodplasma may be only about 0.05 to about 0.1 mM. Oxygen concentrations inother tissue fluids may be even lower. As the chemical reaction, andthus, the sensor signal, is limited by the reactant that is present inthe sensor's reaction zone at the lowest concentration, an implantedsensor of simple construction would remain limited by oxygen, and wouldtherefore be insensitive to the metabolite of interest (e.g. glucose).Thus, there is a need for differential control of the permeability ofthe sensor diffusion device (e.g., “membrane”) to restrict or modulatethe flux of the metabolite of interest (e.g. glucose), and provide astoichiometric equivalent or excess of oxygen in the reaction zone. Thesensor incorporating such a membrane can then be sensitive to themetabolite of interest over the physiologic range. Also, for successfulfunctioning of the implanted sensor, the membrane material exposed tothe bodily tissue must further be biocompatible, or elicit a favorableresponse from the body. Several membrane solutions have been proposed todate.

One such solution has been through the use of macroporous or microporousmembranes to ratio the diffusion of oxygen and glucose to the sensingelements, such as that set forth in U.S. Pat. No. 4,759,828 to Young,which discloses use of a laminated membrane with an outer microporousmembrane having a pore size of 10 to 125 A (Angstrom) to limit thediffusion of glucose molecules. However, one problem with the use of amacroporous or microporous membrane relates to exposure of the sensingelement of the sensor to the environment of the body, which can resultin “fouling” or other deleterious effects. Another solution is disclosedin U.S. Pat. No. 4,671,288 to Gough, which describes a cylindricaldevice, implantable in an artery or vein, which is permeable to glucoseonly at an end of the device, and with both the curved surface and endpermeable to oxygen. In vascular applications, the advantage is directaccess to blood glucose, leading to a relatively rapid response.However, a major disadvantage of vascular implantation is thepossibility of eliciting blood clots or vascular wall damage, as notedsupra.

U.S. Pat. No. 5,660,163 to Schulman discloses another solution throughuse of a silicone rubber membrane containing at least one “pocket”filled with glucose oxidase in a gelatinous glucose and oxygen-permeablematerial located over a first working electrode, such that the length ofthe “pocket” is a multiple of its thickness to optimize the linearitybetween current and the glucose concentration measurement. However,because the long axis of the “pocket” is oriented parallel to theelectrode surface, this design may be less amenable to miniaturizationfor tissue implantation, and may suffer from yet other disabilitiesrelating thereto.

Still further, another solution has been to utilize a composite membranethat is hydrophilic and also contains small hydrophobic domains toincrease the membrane's overall gas solubility, giving rise todifferential permeability of glucose and oxygen (e.g. U.S. Pat. Nos.4,484,987 and 4,890,620 to Gough). However, one salient disadvantage ofthis approach relates to the requirement that the amount of hydrophobicpolymer phase must be relatively large to allow for adequate oxygenpermeability. This substantially reduces the hydrophilic volumeavailable for enzyme inclusion sufficient to counter inactivation duringlong-term operation.

Further, another alternative is described in U.S. Pat. No. 4,650,547 toGough, which discloses a “stratified” structure in which the electrodewas first overlaid with an enzyme-containing layer, and second with anon-glucose-permeable membrane. The resulting structure is permeable tooxygen over a large portion of the surface of the membrane, whereasglucose can only reach the enzyme through the “edge” of the device, thusregulating access of the reactants to the enzyme.

Another significant concern in the context of e.g., implantable solidtissue devices, is the so-called “foreign body response” or FBR, whereinthe host's physiology proximate to the implanted sensor is irritated oradversely stimulated into an antibody-modulated or other response whichcan be deleterious to the operation of the implanted device, especiallyover longer periods of time. For implanted devices that depend ondiffusive transport of solutes to or from the bloodstream (e.g.implanted chemical sensors), such responses can negatively impact deviceoperation due to an increase in mass transfer resistance between thebloodstream and active portions of the device surface resulting from anFBR-mediated development of fibrous tissue surrounding the device. TheFBR also can complicate explants of the implanted device (due to, e.g.,the FBR causing significant encapsulation of the implanted device,thereby increasing its effective size when explanted), and result in yetother disabilities. Thus, accounting for (and minimizing) the FBRremains an important consideration for literally any implanted device.Some prior art solutions for implantable sensors have attempted to uselayers external to the sensing enzyme region to actively modulate oreliminate the FBR. Such approaches have typically used materials forsuch layer(s) which are designed to encourage blood vessel growth andperfusion in the vicinity of the sensor or into the layer(s), which isundesirable, because such modulated responses are often not predictableand furthermore may not be sustainable for extended durations.

Accuracy is also an important consideration for implanted analytesensors, especially in the context of blood glucose monitoring. Hence,ensuring accurate measurement for extended periods of time (andminimizing the need for any other confirmatory or similar analyses) isof great significance.

Similarly, having adequate dynamic range for the implanted sensor isimportant, particularly as it relates to accuracy. Simply stated, theimplanted sensor device should be able to accurately measure the targetanalyte over the entire normal (or even abnormal) range of values thatmay be encountered within the host's physiology.

As such, there is a compelling need for apparatus and methods directedto an implantable analyte (e.g., glucose) sensor designed to overcomethe “oxygen deficit” problem and the disadvantages associated with theprior art devices discussed above (including FBR), while maintaining ahigh degree of accuracy (including over a broad dynamic range), androbustness for extended periods of time. Methods of reliablymanufacturing such sensors are also needed.

SUMMARY

The present disclosure satisfies the foregoing needs by providing, interalia, improved apparatus and associated methods for in vivo glucosemonitoring, including for example within an implantable sensor apparatusor other electronic device within a living subject.

In a first aspect of the disclosure, a sensor apparatus is disclosed. Inone embodiment, the sensor apparatus is configured for (full)implantation in e.g., the solid tissue of a living being, and includesone or more sensor elements capable of measuring blood glucose levelwith a prescribed range of response. In one variant, each sensor elementincludes a hydrophobic outer membrane element, a non-enzymatic membrane,and an enzyme matrix, and the range of response (at a given oxygenlevel) of each sensor element is selectable or controllable throughcontrol of one or more physical attributes of the non-enzymatic membraneand a “spout” structure formed therein (e.g., base height, basediameter, spout height, and/or spout diameter).

In one implementation, the outer membrane is comprised at least in partof a silicone rubber compound, and a single parameter of those listedabove (e.g., the spout diameter) is modified to effect the desired rangeof response.

In one variant of the foregoing implementation, the non-enzymaticmembrane comprises a crosslinked albumin material that is substantiallypermeable to at least glucose and oxygen present at the outer surfacethereof. The non-enzymatic membrane is also placed in contact with a topsurface of the enzyme material so as to permit diffusion of the oxygenand glucose into the enzyme material to permit chemical interactiontherein, while affording the solid tissue of the host an unreactive“buffer zone,” thereby isolating the host tissue from direct contactwith the enzyme material.

In another variant, the response time (at a given oxygen level) of eachsensor element is selectable or controllable through control of one ormore physical attributes of the non-enzyme membrane and/or itssurrounding “spout” structure (e.g., base height, spout height, and/ornon-enzyme membrane thickness).

In one implementation, the outer membrane is comprised at least in partof a silicone rubber compound, the non-enzyme membrane comprises analbumin-based compound, and a single parameter of those listed above(e.g., the spout height) is modified to effect the desired response time(or rate of response).

In another variant, the sensor apparatus includes one or more sensorelements capable of measuring blood glucose level both (i) within aprescribed range of response and (ii) within a prescribed time period.Each sensor element includes a non-enzymatic membrane element (e.g.,including crosslinked albumin), the range of response (at a given oxygenlevel) of the sensor element(s) is selectable or controllable throughcontrol of base diameter and/or spout diameter, and the rate or timingof response (at a given oxygen level) of the sensor element(s) isselectable or controllable through control of base height and/or spoutheight.

In another embodiment, the sensor apparatus includes: signal processingcircuitry and at least one detector element in signal communication withthe signal processing circuitry. The at least one detector elementincludes: a substantially enclosed cavity, the substantially enclosedcavity comprising at least one enzymatic substance, and at least oneaperture in communication with the cavity; an electrolyte layer; atleast one electrode disposed at least partly within or contacting theelectrolyte layer; and a non-enzymatic membrane at least partlyoccluding the aperture, the non-enzymatic membrane comprising a materialat least partly permeable to an analyte yet which does not exacerbate anFBR (including fibrous tissue growth). In one variant, the non-enzymaticmembrane comprises a crosslinked albumin-based material, and the atleast one detector element is configured to utilize chemical interactionbetween at least the analyte and the enzymatic substance to enablegeneration of an electrical signal at the electrode via the electrolytelayer, the electrical signal relating to a concentration of the analytein a region external to the cavity and the membrane.

In a second aspect, a method of forming a sensor element for use on,inter alia, an implantable sensor apparatus is disclosed. In oneembodiment, the method includes forming the aforementioned outerhydrophobic membrane of the sensor element using a molding process, suchthat the desired parameter value(s) is/are achieved (e.g., desiredaperture placement and diameter), and disposing an enzymatic materialwithin an interior cavity of the sensor element, as well as anon-enzymatic tissue-contacting layer over top of the enzymatic materialand within the aperture. In one variant, the outer membrane comprises asilicone rubber compound that can be molded according to the foregoingmethodology, and the non-enzymatic tissue-contacting layer includesalbumin. The albumin layer is in one implementation configured tomitigate excessive FBR.

In a third aspect, a method of configuring a sensor apparatus to achievea desired level of performance is disclosed. In one embodiment, thesensor apparatus includes one or more elements with permeable membranes,and the method includes configuring one or more parameters relating tothe membrane(s) to achieve the desired level of performance. In onevariant, the level of performance relates to a detection range ofresponse of glucose concentration in the blood of a living being, andthe one or more parameters comprise a diameter or an area associatedwith at least a portion of the permeable membrane(s), and/or itsthickness.

In another aspect, a membrane useful with an in vivo sensor apparatus isdisclosed. In one embodiment, the membrane is formed at least in part ofa crosslinked albumin-based compound, and the membrane is specificallyconfigured with respect to at least one parameter thereof (e.g., heightrelative to a base, thickness, and/or diameter, etc.) to produce adesired level of performance or response with respect to one or moresubstances (e.g., glucose in blood), such as rate of permeation ofglucose through the membrane, or detectable range of concentrations ofthe enzyme. In one variant, the albumin-based membrane is configured soas not to exacerbate an FBR after implantation of the host sensorapparatus.

In another aspect of the disclosure, a sensor apparatus withheterogeneous sensor elements is disclosed. In one embodiment, thesensor apparatus is configured for (full) implantation in e.g., thesolid tissue of a living being, and includes two or more sensor elementscapable of measuring blood glucose level in at least one of: (i)different ranges of response, and/or (ii) different times or rates ofresponse. In one variant, each sensor element includes a non-enzymaticmembrane element in contact with the solid tissue, and the range ofresponse (at a given oxygen level) of each sensor element is selectableor controllable through control of one or more physical attributes ofthe membrane and/or its surrounding “spout” structure (e.g., basediameter and and/or spout diameter). The two or more sensor elementsutilize membranes having respective different ranges of response due toe.g., different physical diameters thereof.

In another implementation, the different ranges of response to glucoselevel at least partly overlap one another. In yet anotherimplementation, the different ranges of response to glucose level aresubstantially contiguous, but do not significantly overlap one another.In yet a further implementation, the different ranges of response toglucose level are neither contiguous or overlap one another; i.e., areseparated by one or more ranges (e.g., those not of interest).

In another variant, each sensor element includes a non-enzymaticmembrane element (e.g., an albumin-based membrane such as thatreferenced supra), and the time or rate of response (at a given oxygenand glucose level) of each sensor element is selectable or controllablethrough control of one or more physical attributes of the membraneand/or its surrounding “spout” structure (e.g., membrane thickness,and/or height of the spout above a base region). The two or more sensorelements utilize non-enzyme membranes having respective different ratesof response due to e.g., different physical diameters or otherproperties thereof. In one implementation, eight (8) sensor elements areincluded on the sensor apparatus, two (2) of which are configured tomeasure glucose level within a first range of times, and two of whichare configured to measure glucose level within a second range of times;the remaining four (4) sensor elements are used to measure oxygenconcentration (i.e., are reference elements).

In another variant, combinations of the foregoing are utilized; e.g.,the sensor apparatus includes sensor elements having heterogeneity withrespect to both range of response (concentration) and rate of response(rate of permeation).

In yet a further aspect of the disclosure, a sensor elementconfiguration is disclosed. In one embodiment, the sensor element isused as part of an implantable sensor apparatus, and the configurationincludes an enzyme-free outer layer or portion which prevents tissue ofthe host being from contacting immobilized underlying enzymes (e.g.,glucose oxidase and catalase) within the sensor element when the sensorelement is in vivo, thereby both (i) mitigating or eliminating a foreignbody response (FBR) within the tissue due to lack of exposure to theenzymes or byproducts of the underlying reaction (e.g., peroxide), and(ii) ultimately enhancing operation and longevity of the sensorapparatus due to, inter alia, the aforementioned lack of FBR and the(excess) immobilized enzymes. In one variant, the enzyme-free outerlayer or portion comprises a soluble protein-based (e.g., albumin-based)material which is cured using a cross-linking agent, and which isconfigured so as not to encourage blood vessel growth into the membraneafter implantation of the sensor element into a living host.

In a further aspect of the disclosure, a method of mitigating foreignbody response (FBR) associated with an implanted analyte detectorelement is disclosed. In one embodiment, the method includes creating anenzyme-free buffer zone between a reactive enzymatic portion of thedetector element using a non-enzymatic membrane or layer that ispermeable to both the analyte (e.g., glucose) and oxygen. In onevariant, the enzymatic portion comprises a mechanically stable matrix,such that the enzymes of the matrix are substantially immobilized and donot migrate outward toward the host's tissue through the non-enzymaticmembrane or layer, but the analyte and oxygen from the tissue canmigrate inward. In one implementation, the non-enzymatic membranematerial is selected so as to not exacerbate the FBR, thereby mitigatingfibrous tissue formation and making subsequent explants of the detectorelement easier due to limited FBR-induced encapsulation.

In another aspect, a method of controlling at least one operatingcharacteristic of an oxygen-based blood analyte sensing element isdisclosed. In one embodiment, the sensing element has an enzyme-filledcavity with a base region, the cavity accessible via an aperture and anat least partly protein-based permeable membrane covering the aperture,and the method includes controlling at least one of a dimension of theaperture or the base so as to selectively control the diffusion ofanalyte and oxygen into the cavity, thereby causing the sensing elementto exhibit the desired operating characteristic when operated. In onevariant, the analyte comprises glucose; the enzyme filling comprisesglucose oxidase and catalase; and the desired operating characteristiccomprises a range of analyte concentration that can be detected by thesensing element, and the at least one dimension comprises at least oneof: (i) a diameter of the aperture, and/or (ii) a diameter of the baseregion.

In another variant, the analyte comprises glucose; the enzyme fillingcomprises glucose oxidase and catalase; and the desired operatingcharacteristic comprises a time within which an analyte concentrationcan be detected by the sensing element, and the at least one dimensioncomprises at least one of: (i) a height of the aperture relative to aheight of the base region.

In yet a further aspect of the disclosure, a method of operating anoxygen-based blood analyte sensing element is described. In oneembodiment, the element includes an enzyme-filled cavity with a baseregion, the cavity accessible via an aperture and an at least partlyenzyme-free permeable membrane covering the aperture, and the methodincludes: selectively controlling the migration of at least an analyteinto the cavity through the enzyme-free membrane; creating residualoxygen within the cavity; and selectively controlling the migration ofat least portions of the residual oxygen to at least one electrode ofthe sensing element. In one variant, the selective control of themigration of analyte and the selective control of the migration ofoxygen cooperate to cause the sensing element to exhibit a desiredoperating characteristic.

In one implementation, the desired operating characteristic comprises arange of glucose concentrations detectable by the sensing element, andthe selectively controlling the migration of the glucose is accomplishedat least in part by selectively choosing an area of the aperture.

In still another aspect, an analyte detection apparatus is disclosed. Inone embodiment, the apparatus includes: a substrate; at least oneelectrode disposed on or within the substrate, the at least oneelectrode comprising at least a terminal configured to enable electricalsignals to be communicated from the electrode to a circuit; anelectrolyte material in communication with at least a portion of the atleast one electrode; a first membrane element in contact with at least aportion of the electrolyte material; a second membrane elementcomprising a cavity formed therein, and at least one aperture; anenzymatic material disposed within the cavity and configured to interactwith the analyte and at least a portion of the oxygen entering thecavity via the aperture; and a non-enzymatic material configured tosubstantially occlude the aperture and frustrate migration of anyenzymes from the enzymatic material outward to the tissue of the livinghost, yet permit diffusion of analyte and oxygen therethrough. In onevariant, a response characteristic of the analyte detection apparatus iscontrolled at least in part by a shape of the second membrane element.

In another variant, the substrate comprises a ceramic material, thefirst membrane comprises a polymeric material, and the second membraneelement comprises a silicone rubber-based material; the analytecomprises glucose, and the enzymatic material comprises glucose oxidaseand catalase disposed in a crosslinked matrix; and the non-enzymaticmaterial comprises albumin.

In a further aspect of the disclosure, a dynamically variable sensorapparatus is disclosed. In one embodiment, the sensor apparatusincludes: signal processing circuitry; and at least one first detectorelement and at least one second detector element each in signalcommunication with the signal processing circuitry. In one variant, theat least one first and second detector elements each comprise: a partlyenclosed cavity, the cavity comprising at least one enzymatic substance,and at least one aperture in communication with the cavity, the apertureat least partly obscured with a non-enzyme yet permeable substance; anelectrolyte layer; and at least one electrode disposed at least partlywithin or contacting the electrolyte layer; and the at least one firstand second detector elements are each configured to utilize chemicalinteraction between at least the analyte and their respective enzymaticsubstance to enable generation of an electrical signal at theirrespective electrodes via their respective electrolyte layer, theelectrical signal relating to a concentration of the analyte in a regionexternal to their respective cavities.

In one implementation, at least one of (i) a shape or dimension of theaperture, (ii) a thickness of the non-enzyme yet permeable substance,and/or (iii) a shape or dimension of the cavity, for the at least onefirst detector element is different from that for the second at leastone detector element, thereby enabling the at least one first detectorelement and the at least one second detector element to have a differentoperational characteristic from the other.

In yet another aspect of the disclosure, a method of mitigating aforeign body response (FBR) within a living being while monitoring bloodglucose level using an implanted sensor apparatus is described. In oneembodiment, the method includes: allowing at least oxygen molecules andglucose molecules from the living being's blood to permeate through anon-enzymatic layer or membrane to an enzyme-containing material withwhich the oxygen molecules and glucose molecules can chemicallyinteract, the chemical interaction enabling the monitoring; and at leastmitigating egress of enzymes within the enzyme-containing materialoutward through the layer or membrane. In one variant, the non-enzymaticmaterial comprises a protein-based (e.g., albumin) substance which ischemically crosslinked, and which does not encourage blood vessel growthinto its thickness.

In a further aspect, a method of manufacturing an analyte detectorelement is described. In one embodiment, the method includes: providinga substrate; embedding at least one electrode within the substrate;disposing an electrolyte over at least a portion of the substrate,including at least a portion of the electrode; forming an inner membraneover at least a portion of the electrolyte; forming an outer membranehaving an aperture formed therein, and disposing the outer membrane overthe inner membrane such that the inner and outer membranes form acavity; disposing an enzyme matrix material into the cavity such that itat least contacts the inner membrane and aperture; and forming a layerof non-enzymatic material within the aperture such that it at leastpartly contacts the enzyme matrix material.

In one variant of the method, the outer and inner membranes comprise anelastomeric material (e.g., silicone rubber), and the outer membrane isadhered (e.g., via a room temperature vulcanizing (RTV) adhesive) to theinner membrane in order to create the cavity. The enzyme matrix is thenintroduced into the cavity via the aperture; e.g., the matrix issubstantially liquefied, and is poured or piped into the cavity so as tosubstantially fill the cavity, and exclude air therefrom, while alsomaintaining the aperture region entirely enzyme-free. The enzyme matrixis then cured (e.g., chemically crosslinked), and subsequently thenon-enzymatic material (also in a substantially liquefied or flowableform) is poured or piped into the top portion of the cavity via theaperture (i.e., atop the cured enzyme matrix), and subsequentlychemically crosslinked as well.

In yet another aspect of the disclosure, a method of manufacturing ananalyte detector element is disclosed. In one embodiment, the methodincludes: disposing an electrolyte material over at least a portion ofan active surface of an electrode; forming a hydrophobic membranestructure over the electrolyte material, the hydrophobic membranestructure comprising at least (i) a cavity and (ii) a spout region incommunication with the cavity; forming an enzyme matrix within thecavity such that the enzyme matrix contacts at least a portion of abottom interior wall of the cavity; and forming a hydrophilic membraneat least partly within the spout region such that the hydrophilicmembrane contacts at least a portion of a top surface of the enzymematrix.

In one variant of the method, the forming of the spout region at leastin part comprises forming a specified three-dimensional shape of thespout region, the specified three-dimensional shape configured toregulate analyte diffusion across the hydrophilic membrane.

In another variant, the forming of the enzyme matrix at least in partcomprises: disposing a flowable enzyme matrix material within thecavity, the flowable enzyme matrix material at least in part comprisinga first chemically cross-linkable material; applying a first chemicalcrosslinking agent to the flowable enzyme matrix material within thecavity, thereby causing: cross-linking of the first chemicallycross-linkable material, and forming of a cross-linked enzyme matrixdisposed within the cavity and at least partly coincident with a bottomedge of an inner wall of the spout region.

In one implementation, the forming of the hydrophilic membrane at leastin part comprises: subsequent to the applying of the first chemicalcrosslinking agent and the forming of the cross-linked enzyme matrix,disposing a flowable hydrophilic membrane material at least partiallywithin the spout region, the flowable hydrophilic membrane material atleast in part comprising a second chemically cross-linkable material;and applying a second chemical cross-linking agent, the second chemicalcrosslinking agent causing: cross-linking of the second chemicallycross-linkable material; forming of a cross-linked hydrophilic membranedisposed within the spout region and substantially coincident with a topedge of an outer wall of the spout region; and bonding of a bottomsurface of the cross-linked hydrophilic membrane to a top surface of thecross-linked enzyme matrix.

Other features and advantages of the present disclosure will immediatelybe recognized by persons of ordinary skill in the art with reference tothe attached drawings and detailed description of exemplary embodimentsas given below.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a front perspective view of one exemplary embodiment of afully implantable sensor apparatus according to the present disclosure.

FIGS. 1A-1C are top, bottom, and side elevation views, respectively, ofthe exemplary sensor apparatus of FIG. 1.

FIG. 2 is a side cross-sectional view of one exemplary detector elementof a detector array in a fully implantable sensor apparatus according tothe present disclosure.

FIG. 2A is a side cross-sectional view of one exemplary spout region(outer non-enzyme membrane removed) of a detector element of a detectorarray in a fully implantable sensor apparatus according to oneembodiment of the present disclosure.

FIG. 2B is a graphical representation of top elevation views of avariety of different exemplary embodiments of sensor membrane aperturesaccording to the present disclosure.

FIGS. 2C-2F are side cross-sectional views of respective ones of avariety of different exemplary embodiments of sensor membrane aperturesaccording to the present disclosure.

FIG. 3 is a top elevation view of another exemplary embodiment of thesensor apparatus of present disclosure, wherein multiplesensor/reference pairs with at least partly differing glucose sensingranges are used on a common device.

FIG. 4 is a logical flow diagram of one embodiment of a method ofmanufacturing a sensor element according to the present disclosure.

All Figures © Copyright 2016 GlySens Incorporated. All rights reserved.

DETAILED DESCRIPTION

Reference is now made to the drawings, wherein like numerals refer tolike parts throughout.

Detailed Description of Exemplary Embodiments

Exemplary embodiments of the present disclosure are now described indetail. While these embodiments are primarily discussed in the contextof a fully implantable glucose sensor, such as those exemplaryembodiments described herein, and/or those set forth in U.S. PatentApplication Publication No. 2013/0197332 filed Jul. 26, 2012 entitled“Tissue Implantable Sensor With Hermetically Sealed Housing;” U.S. Pat.No. 7,894,870 to Lucisano et al. issued Feb. 22, 2011 and entitled“Hermetic implantable sensor;” U.S. Patent Application Publication No.20110137142 to Lucisano et al. published Jun. 9, 2011 and entitled“Hermetic Implantable Sensor;” U.S. Pat. No. 8,763,245 to Lucisano etal. issued Jul. 1, 2014 and entitled “Hermetic feedthrough assembly forceramic body;” U.S. Patent Application Publication No. 20140309510 toLucisano et al. published Oct. 16, 2014 and entitled “HermeticFeedthrough Assembly for Ceramic Body;” U.S. Pat. No. 7,248,912 toGough, et al. issued Jul. 24, 2007 and entitled “Tissue implantablesensors for measurement of blood solutes;” and U.S. Pat. No. 7,871,456to Gough et al. issued Jan. 18, 2011 and entitled “Membranes withcontrolled permeability to polar and apolar molecules in solution andmethods of making same,” each of the foregoing incorporated herein byreference in its entirety, it will be recognized by those of ordinaryskill that the present disclosure is not so limited. In fact, thevarious aspects of the disclosure are useful with, inter alia, othertypes of implantable sensors and/or electronic devices.

Further, while the following embodiments describe specificimplementations of e.g., oxygen-based multi-sensor element devices, andhaving specific configurations, those of ordinary skill in the relatedarts will readily appreciate that such descriptions are purelyillustrative, and in fact the methods and apparatus described herein canbe used consistent with, and without limitation: (i) in living beingsother than humans; (ii) other types or configurations of sensors (e.g.,peroxide-based glucose sensors, or sensors other than glucose sensors,such as e.g., for other analytes such as urea, lactate); and/or (iii)devices intended to deliver substances to the body (e.g. implanted drugpumps, drug-eluting solid materials, and encapsulated cell-basedimplants, etc.); and/or other devices (e.g., non-sensors andnon-substance delivery devices).

As used herein, the terms “detector” and “sensor” refer withoutlimitation to a device that generates, or can be made to generate, asignal indicative of a measured parameter, such as the concentration ofan analyte (e.g., glucose or oxygen). Such a device may be based onelectrochemical, electrical, optical, mechanical, thermal, or otherprinciples as generally known in the art. Such a device may consist ofone or more components, including for example, one, two, three, or fourelectrodes, and may further incorporate immobilized enzymes or otherbiological or physical components, such as membranes, to provide orenhance sensitivity or specificity for the analyte.

As used herein the term “membrane” refers without limitation to asubstance, layer or element configured to have at least one desiredproperty relative to the aforementioned analyte, such as e.g., apermeability to a given type of analyte or sub stance.

As used herein, the terms “enzyme free” and “non-enzymatic” include,without limitation, materials that are completely enzyme-free, andmaterials that are substantially enzyme free (e.g., may have a smallpercentage of residual or unintentional enzymes).

Likewise, as used herein, the terms “top,” “bottom,” “side,” “up,”“down,” and the like merely connote, without limitation, a relativeposition or geometry of one component to another, and in no way connotean absolute frame of reference or any required orientation. For example,a “top” portion of a component may actually reside below a “bottom”portion when the component is mounted to another device (e.g., hostsensor).

Overview

In one exemplary aspect, the present disclosure provides improvedenzymatic detectors and associated membrane apparatus, and associatedmethods of manufacturing and use, such as within a fully implantablesensor apparatus of the type described in U.S. patent application Ser.No. 14/982,346 entitled “Implantable Sensor Apparatus and Methods,”previously incorporated by reference herein. Advantageously, theapparatus and methods of the disclosure enable, inter alia,substantially continuous, long-term and accurate monitoring of bloodglucose levels in living beings using the aforementioned implantablesensor apparatus, without the need for prior art “finger sticks,”transcutaneous apparatus worn on external surfaces of the body, orintravenous devices, each having their own disabilities as previouslydescribed.

Specifically, the present disclosure describes a novel sensor (detector)element configuration, including use of selectively configured membraneelements and enzyme region shapes, which enable accurate detection ofblood glucose level within the solid tissue of the host for extendedperiods of time, and within desired ranges and/or rates of response. Theperformance of the detector elements may be controlled through variationof one or more physical parameters of the membrane elements (e.g.,dimensions, shapes, etc.), including an access or “spout” region, so asto allow for precise measurement of the target analyte, while alsomaintaining isolation between the host's tissue and the underlyingenzymes and also potentially between the host tissue and reactionbyproducts used in the sensor element, thereby advantageously minimizingforeign body response (FBR) to the device while implanted, particularlyin the region(s) where sensing of the target analyte is performed.

In one approach, an enzyme-free layer or membrane is formed within thespout region and used in conjunction with a substantially immobilizedunderlying enzyme material, such that the analyte (e.g., glucosemolecules) and oxygen molecules can permeate through to the enzymeregion, but the enzymes (e.g., catalase and glucose oxidase) do notpermeate outward. The spout region of the sensor element is alsoadvantageously maintained substantially enzyme-free duringmanufacturing, and a reasonably tight “seal” is formed around aperiphery of the enzyme-free layer or membrane after curing (and theenzyme-free layer may even be bonded to its surrounding structure), suchthat undesired enzyme migration or contact with the living host's tissueare avoided.

The disclosed configuration (including use of the enzyme-free layer)advantageously does not encourage blood vessel ingrowth, which interalia, enables accurate sensor apparatus operation during periods ofextended implantation. By not encouraging such ingrowth, otherwiseunstable/unpredictable modulation of FBR to the extent required toencourage blood vessel ingrowth is wholly avoided. In one variant, suchingrowth is frustrated (or not encouraged) through selective control ofpore sizes within the non-enzymatic layer.

Control of response range and/or rate also permits easy “customization”of sensor elements, whether on a per-element or per-sensor apparatusbasis. For example, the techniques of the present disclosure allow forready construction of an implantable sensor apparatus having multipleheterogeneous detector elements with respective multiple ranges ofsensitivity and/or rates of detection, thereby extending the dynamicrange of the sensor apparatus (both in terms of analyte concentrationand/or time, as desired).

Moreover, in one variant, the various heterogeneous detector elementscan be selectively switched on/off (even while the sensor apparatus isin vivo), so as to, e.g., accommodate “on the fly” changes to bloodglucose concentration occurring within the host, or to maintain thedetector elements within a known or desirable range of accuracy orsensitivity.

Methods of manufacturing the aforementioned membranes and sensorelements are also disclosed herein.

Exemplary Implantable Sensor Apparatus

Referring now to FIGS. 1-1C, one exemplary embodiment of a sensorapparatus useful with various aspects of the present disclosure is shownand described.

As shown in FIGS. 1-1C, the exemplary sensor apparatus 100 comprises asomewhat planar housing structure 102 with a sensing region 104 disposedon one side thereof (i.e., a top face 102 a). The exemplarysubstantially planar shape of the housing 102 provides mechanicalstability for the sensor apparatus 100 after implantation, therebyhelping to preserve the orientation of the apparatus 100 (e.g., withsensing region 104 facing away from the epidermis and toward theproximate fascial layer), resisting rotation around its longitudinalaxis 108, and translation, or rotation about its transverse axis 110,which might otherwise be caused by e.g., normal patient ambulation ormotion, sudden accelerations or decelerations (due to e.g., automobileaccidents, operation of high-performance vehicles such as aircraft), orother events or conditions. Notwithstanding, the present disclosurecontemplates sensor apparatus of shapes and/or sizes other than that ofthe exemplary apparatus 100, including use of means to maintain thedesired orientation and position such as e.g., protruding tabs,“anchoring” the sensor apparatus to surrounding physiological structuressuch as the fascial layer by means of sutures or the like, attachedtissue ingrowth structures, and so forth.

The exemplary sensor apparatus of FIGS. 1-1C further includes a detectorarray 104 comprising a plurality of individual detectors elements 106 onthe top face 102 a of the sensor apparatus housing 100. In exemplaryembodiments, the detector array 104 of the present invention includeseight (8) detector elements 106 with associated membranes (not shown)disposed on a detector substrate 112 (e.g. ceramic disk), which functionas a group. As will be discussed in greater detail infra, the detectorelements 106 in the illustrated embodiment comprise a plurality ofprimary detectors including associated membranes containing glucoseoxidase and catalase to measure glucose-dependent oxygen levels, and aplurality of secondary detectors including associated membranes withoutbonded enzymes to measure background oxygen levels.

In operation, a signal-processing element (not shown) measures thecurrent difference between the glucose-dependent oxygen current and thebackground oxygen current, to produce a glucose-dependent differencecurrent. As such, the exemplary sensor apparatus 100 of the presentdisclosure utilizes an “oxygen-sensing differential measurement,” bycomparison of the glucose-dependent oxygen signal to the backgroundoxygen signal that produces, upon further signal processing, acontinuous real-time blood glucose concentration measurement. It will beappreciated, however, that the methods and apparatus described hereinare in no way limited to such “differential” schemes.

The exemplary sensor apparatus of FIGS. 1-1C also includes a plurality(three in this instance) of tabs or anchor apparatus 113 disposedsubstantially peripheral on the apparatus housing. These anchorapparatus provide the implanting surgeon with the opportunity to anchorthe apparatus to the anatomy of the living subject, so as to frustratetranslation and/or rotation of the sensor apparatus 200 within thesubject immediately after implantation but before any body response(e.g., FBR) of the subject has a chance to immobilize (such as viaencapsulation) the sensor apparatus. See, e.g., the exemplary apparatusand techniques described in co-owned and co-pending U.S. patentapplication Ser. No. 14/982,346 filed Dec. 29, 2015 and entitled“Implantable Sensor Apparatus and Methods,” incorporated herein byreference in its entirety.

In the exemplary configuration, four (4) of each type of detectorelements 106 are included (i.e., four primary, and 4 secondary),although it will be appreciated that any number of detector elements canbe used consistent with the disclosure (including in pairs, or where oneprimary or secondary detector is used in conjunction with two or more ofthe other type of detector). Numerous permutations are possible, allconsidered within the scope of the present disclosure.

Further considerations relating to the total number of detector elements106 on the detector array 104 include, e.g., available surface area ofthe detector substrate 112, which in turn is dictated by a desire tominimize the overall size of the sensor for implantation. In theexemplary embodiments of the sensor apparatus 100, use of a plurality ofdetector elements 106 is important to: 1) maximize the probability thatseveral detectors will be positioned very near an active vascular bed;2) afford the possibility of ignoring a given detector element if itoperates inaccurately, erratically, or becomes nonresponsive over time;and 3) minimize the effects of local variations in analyteconcentration, as well as local variations in the magnitude of anyconfounding phenomena occurring proximate the detector elements (see,inter alia, U.S. Pat. No. 7,248,912 previously incorporated herein, fora discussion of various confounding phenomena).

The exemplary detector elements 106 are of the enzyme-electrode type(some utilizing membranes containing immobilized glucose oxidase andcatalase), such as those exemplary embodiments described herein, and/orthose set forth in U.S. Pat. No. 4,484,987 to Gough, entitled “MethodAnd Membrane Applicable To Implantable Sensor;” U.S. Pat. No. 4,671,288to Gough, entitled “Electrochemical Cell Sensor For ContinuousShort-Term Use In Tissues And Blood;” U.S. Pat. No. 4,650,547 to Gough,entitled “Method And Membrane Applicable To Implantable Sensor;” U.S.Pat. No. 4,890,620 to Gough, entitled “Two-Dimensional Diffusion GlucoseSubstrate Sensing Electrode;” U.S. Pat. No. 5,322,063 to Allen et al.entitled “Hydrophilic Polyurethane Membranes For Electrochemical GlucoseSensors;” U.S. Pat. No. 5,660,163 to Schulman et al., entitled “GlucoseSensor Assembly,” and U.S. Pat. No. 6,721,587 to Gough, entitled“Membrane and Electrode Structure For Implantable Sensor,” each of whichare incorporated herein by reference in its entirety. It will beappreciated, however, that the type and operation of the sensorapparatus 100 may vary; i.e., other types of detector elements/sensorapparatus, configurations, and signal processing techniques thereof maybe used consistent with the various aspects of the present disclosure,including, for example, signal processing techniques based on variouscombinations of signals from individual elements in the otherwisespatially-defined sensing elements pairs.

Methods for calculating the levels of glucose present based on aspecific enzymatic reaction are well known in the art, as are certaincalibration techniques (see, e.g., Choleau, et al., Biosens.Bioelectron., 17:647-654 (2002) and Choleau, et al., Biosens.Bioelectron., 17:641-646 (2002), the teachings of which are incorporatedherein by reference in their entirety). Benchmark data for evaluation ofsensor performance are also available (Bremer, et al, Diabetes Technol.Ther., 3:409-418 (2001), the teachings of which are incorporated hereinby reference).

The exemplary detector elements 106 utilize the following two-stepchemical reaction catalyzed by glucose oxidase and catalase to detectglucose, as described in Armour et al. (Diabetes 39, 1519-1526 (1990)):

Glucose+O₂+H₂O

Gluconate+H₂O₂ (Glucose Oxidase)

H₂O₂

½O₂+H₂O (Catalase)

Glucose+½O₂

Gluconate (Net)

In the exemplary embodiment of the (primary) detector elements describedherein, the two enzymes (glucose oxidase and catalase) are entrapped ormore preferably immobilized within a gel matrix (discussed infra) thatis e.g., crosslinked for mechanical and chemical stability, and which isin operative contact with a working electrode to electrochemicallydetect oxygen. It will be appreciated that while a chemical or othercrosslinking technique can be used consistent with the disclosure toimmobilize the enzymes, other approaches for immobilization and/ormechanical stabilization of the enzyme matrix (primary detectors) and/ornon-enzyme matrix (secondary detectors) may also be used, whether aloneor in combination with the foregoing. For instance, in one variant, anon-cross-linked (yet mechanically stable) gel could be used. As anotheralternative, a porifera or other sponge/sponge-like structure could beused (e.g., with the enzyme disposed within the pores or ostia of thestructure for mechanical stability). In yet another approach, a ladder,scaffold, or three-dimensional mesh structure could be used to supportthe enzymatic material.

In the illustrated embodiment, glucose and ambient oxygen diffuse intothe gel matrix and encounter the enzymes, the above reactions occur, andthe oxygen that is not consumed in the process is detected by theworking or primary electrode.

Exemplary Detector Element with Associated Membranes

As shown in FIG. 2, an exemplary individual (primary) detector element106 according to the present disclosure is shown associated withdetector substrate 214 (e.g. a ceramic substrate), and generallycomprises a plurality of membranes and/or layers, including e.g., theinsulating layer 260, and electrolyte layer 250, an enzymatic gel matrixof the type described above 240, an inner membrane 220, an exteriormembrane shell 230, and a non-enzymatic membrane 277. Such membranes andlayers are associated with the structure of individual detectorelements, although certain membrane layers can be disposed in acontinuous fashion across the entire detector array surface or portionsthereof that include multiple detectors, such as for economies of scale(e.g., when multiple detectors are fabricated simultaneously), or formaintaining consistency between the individual detector elements byvirtue of making their constituent components as identical as possible.

As noted above, the exemplary sensor apparatus 100 includes both primary(enzymatic) and secondary (non-enzymatic) detector elements; while FIG.2 illustrates one embodiment of the former, the latter is in theexemplary embodiment generally similar in structure, with the exceptionthat the enzymatic matrix 240 is replaced with a non-enzymatic matrix orother structure; i.e., in the case of a glucose-sensitive primarydetector, it does not include the glucose oxidase and catalase describedelsewhere herein. It will be appreciated, however, that the constructionof primary and secondary detector elements need not be substantiallysimilar, and in fact may differ significantly in construction so long asthe desired performance attributes are maintained.

The detector element 106 further comprises a working electrode 217 inoperative contact by means of electrolyte layer 250 with a counterelectrode 219 and a reference electrode 218, and their associatedfeedthroughs 280 (details of the exemplary feedthroughs 280 aredescribed in U.S. Pat. No. 8,763,245 to Lucisano et al. entitled“Hermetic feedthrough assembly for ceramic body,” previouslyincorporated by reference herein). The working electrode 217 comprisesan oxygen-detecting catalytic surface producing a glucose-modulated,oxygen-dependent current (discussed infra), reference electrode 218comprises an electrochemical potential reference contact to electrolytelayer 250, and counter electrode 219 is operably connected by means ofelectrolyte layer 250 to the working electrode 217 and referenceelectrode 218. An electrical potentiostat circuit (not shown) is coupledto the electrodes 217, 218, and 219 to maintain a fixed potentialbetween the working and reference electrode by passing current betweenthe working and counter electrodes while preferably maintaining thereference electrode at high impedance. Such potentiostat circuitry iswell known in the art (for an example, see U.S. Pat. No. 4,703,756 toGough et al. entitled “Complete glucose monitoring system with animplantable, telemetered sensor module,” incorporated herein byreference in its entirety).

Generally, the thickness of each of the membranes disclosed herein isnot particularly limited, as long as the desired permeability propertiesare achieved. However, particular requirements for sensor response time,glucose concentration detection range, and/or reduction of antibodyresponse (e.g., FBR), may impose limits on the allowable membranethickness. Membrane thickness can be, for example, about 1 micron toabout 1000 microns, or more particularly, about 10 microns to about 500microns, or more particularly about 25 microns to about 250 microns, ormore particularly about 25 microns to about 75 microns. Very thinmembrane layers, particularly those less than about 10 microns, mayrequire mechanical support to be provided in the form of a backingmembrane, which may be a porous, relatively inert structure. U.S. Pat.No. 7,336,984 and entitled “Membrane and Electrode Structure forImplantable Sensor,” previously incorporated herein, describes exemplarymembrane apparatus, thickness values, and computerized modelingtechniques useful with the various aspects of the present disclosure,although it will be recognized that other techniques, apparatus, andmethods for membrane configuration may be used consistent with thepresent disclosure. The electrolyte layer 250 comprises, in theillustrated embodiment, a layer of hydrophilic electrolyte materialwhich is in direct contact with the working electrode(s) 217, referenceelectrode(s) 218 and counter electrode(s) 219. In variousimplementations, materials for constructing the hydrophilic electrolytelayer 250 include salt-containing gels comprising polyacrylamide,poly(ethylene oxide) poly(hydroxyethylmethacrylate) and its derivatives,and other hydrophilic polymers and copolymers, in both crosslinked andnon-crosslinked form. Various other construction details of theexemplary electrolyte layer 250 are described in U.S. Patent ApplicationPublication No. 2013/0197332 filed Jul. 26, 2012 entitled “TissueImplantable Sensor With Hermetically Sealed Housing,” each incorporatedby reference herein in its entirety.

In an exemplary embodiment, the enzymatic material 240 comprises acrosslinked gel of hydrophilic material including enzymes (e.g., glucoseoxidase and catalase) immobilized within the gel matrix, including abuffer agent and small quantities of a chemical cross-linking agent. Thehydrophilic material is permeable to both a large molecule component(e.g. glucose) and a small molecule component (e.g. oxygen). In variousembodiments, specific materials useful for preparing the enzymaticmaterial 240, include, in addition to an enzyme component,polyacrylamide gels, glutaraldehyde-crosslinked collagen or albumin,poly(hydroxyethylmethacrylate) and its derivatives, and otherhydrophilic polymers and copolymers, in combination with the desiredenzyme or enzymes. The enzymatic material 240 can similarly beconstructed by crosslinking glucose oxidase or other enzymes withchemical crosslinking reagents, without incorporating additionalpolymers.

The enzymatic material 240 is in operative contact with the workingelectrode 217 through the inner membrane 220 and the electrolyte layer250 to allow for the electrochemical detection of oxygen at the workingelectrode 217 modulated by the two-step chemical reaction catalyzed byglucose oxidase and catalase discussed above. To that end, as glucoseand ambient oxygen diffuse into the enzymatic material 240, theyencounter the resident enzymes (glucose oxidase and catalase) and reacttherewith; the oxygen that is not consumed in the reaction(s) diffusesthrough the inner membrane 220 and is detected at the working electrode217 to yield a glucose-dependent oxygen signal.

A hydrophobic material is utilized for inner membrane 220, which isshown in FIG. 2 as being disposed over the electrolyte layer 250. Thehydrophobic material is impermeable to the larger or less solublemolecule component (e.g. glucose) but permeable to the smaller or moresoluble molecule component (e.g. oxygen). In various embodiments,materials useful for preparing hydrophobic layers, including innermembrane 220, as well as membrane shell 230, include organosiliconpolymers, such as polydimethylsiloxane (PDMS) and derivatives thereof,polymers of tetrafluoroethylene, ethylene tetrafluoroethylene, orfluorochloro analogs alone or as copolymers with ethylene or propylene,polyethylene, polypropylene, cellulose acetate, and otheroxygen-permeable polymeric materials.

The inner membrane 220 can also be a continuous layer across the entiredetector array surface, and thus be a single common layer utilized byall detectors in the detector array. It is noted that the inner membrane220, inter alia, protects the working electrode 217, reference electrode218 and counter electrode 219 from drift in sensitivity due to contactwith certain confounding phenomena (e.g. electrode “poisoning”), but theworking electrode 217 will nonetheless be arranged sufficiently close tothe enzymatic material to enable detection of oxygen levels therein.

The (hydrophobic) outer membrane shell 230 is disposed over at least aportion of the enzymatic material 240 (forming a cavity 271 within whichthe material 240 is contained), and is further configured to include anaperture within a “spout” region 270 (discussed in greater detailinfra). In the exemplary embodiment, the membrane shell 230 isseparately provided and adhesively bonded to the inner membrane 220,although it is also contemplated that the inner membrane 220 and themembrane shell 230 can be coextensive and therefore be disposed as onecontinuous membrane layer in which outer membrane shell 230 and innermembrane 220 are of the same uniform thickness of membrane across theindividual detector and array.

As shown in the exemplary embodiment of FIG. 2, inner membrane 220 andmembrane shell 230 are disposed in a manner that creates discretethree-dimensional regions having different thicknesses on the detectorsubstrate 214, which can be utilized to create tissue anti-migrationelements used to achieve stability of location and prevention of devicemigration away from its original implant location. Alternatively, thehydrophobic component may be dispersed as small domains in a continuousphase of the hydrophilic material. Various other construction details ofthe hydrophobic component dispersed as small domains in a continuousphase of hydrophilic material are described in U.S. Pat. Nos. 4,484,987and 4,890,620, each incorporated herein by reference in its entirety.

The exemplary sensor apparatus is made biocompatible to allow for longterm implantation into biological tissue. Thus all membrane structuresthat are in direct contact with the surrounding biological material arebiocompatible and not problematically immunogenic. The membranematerials disclosed herein that are in direct contact with tissue (i.e.,the non-enzymatic membrane element 277 and the outer membrane (shell)230 are generally known to be biocompatible and suitable for long termimplantation. However, in some embodiments, all or discrete regions ofthe sensor may include one or more additional coatings or membranelayers of non-erodible biocompatible material, which may be included toensure that the immunogenic potential of all exposed materials remainssuitably low.

Exemplary Spout Region of the Detector Element

As shown in FIGS. 2-2A, a single spout region 270 of the detectorelement 106 forms a small opening or aperture 276 through the membraneshell 230 to constrain the available surface area of hydrophilicenzymatic material 240 exposed for diffusionally accepting the solute ofinterest (e.g. glucose) from solution. Alternatively, it is contemplatedthat one or more spout regions (and or apertures within a spout region)can exist per detector element.

The shape and dimension of spout region 270 aids in controlling the rateof entry of the solute of interest (e.g. glucose) into enzymaticmaterial 240, and thus impacts the effective operational permeabilityratio of the enzymatic material 240. Such permeability ratio can beexpressed as the maximum detectable ratio of glucose to oxygenconcentration of an enzymatic glucose sensor, where such a sensor isbased on the detection of oxygen unconsumed by the enzyme reaction, andafter taking into account the effects of external mass transferconditions and the enzyme reaction stoichiometry. Detailed discussionsof the relationship between membrane permeability ratio and the maximumdetectable ratio of glucose to oxygen concentration of oxygen-detecting,enzymatic, membrane-based sensors are provided in “Model of aTwo-Substrate Enzyme Electrode for Glucose,” J. K. Leypoldt and D. A.Gough, Analytical Chemistry, 56, 2896 (1984) and “Diffusion and theLimiting Substrate in Two-Substrate Immobilized Enzyme Systems,” J. K.Leypoldt and D. A. Gough, Biotechnology and Bioengineering, XXIV, 2705(1982), incorporated herein by reference. The membranes of the exemplarydetector element described herein are characterized by a permeabilityratio of oxygen to glucose of about 200 to about 1 in units of (mg/dlglucose) per (mmHg oxygen). Note that while this measure of permeabilityratio utilizes units of a glucose concentration to an oxygenconcentration, it is nevertheless a measure of the ratio of oxygen toglucose permeability of the membrane.

The exemplary spout 270 is formed out of the hydrophobic material of themembrane shell 230 without bonded enzymes (e.g., silicone rubber) andadvantageously includes a non-enzymatic outer layer or membrane 277 to,inter alia, prevent direct contact of the immobilized enzymes in theenzymatic material 240 with the surrounding tissue, thereby eliminatingand/or reducing antibody response (e.g., FBR), encapsulation, and/orother deleterious factors. In exemplary embodiments, the non-enzymaticmembrane 277 is further constructed (i.e., with a substantially planarcrosslinked biocompatible matrix possessing pores substantially smallerthan those required to accommodate blood vessel ingrowth, but largeenough to accommodate diffusion of solutes of interest) so as tofrustrate or mitigate blood vessel formation therein. (Suitable poresinclude those with an effective diameter ranging from approximately 10angstroms up to approximately 10 microns.) Herein lies a salient featureof the sensor element of the exemplary embodiment; i.e., the combinationof (i) an enzyme-free biocompatible outer membrane 277, (ii) maintenanceof the spout region substantially free of enzyme material duringmanufacture (see discussion of manufacturing methods below), (iii) useof a non-porous crosslinked structure for the membrane 277, and (iv) useof a biocompatible material (e.g., silicone rubber) for the outermembrane shell 230, dramatically reduces the level of FBR of the hostwhile the device is implanted, thereby allowing for both longerimplantation (due to, inter alia, the reduced level of FBR notinterfering with sensor operation) and easier explants of the device, ascompared to e.g., peroxide-based sensors without such features. Theinner hydrophobic membrane 220 further provides additional insulation ofthe host tissue in the region of the detector 106 against any electricalpotentials which may be present with in the sensor element, therebyfurther aiding in mitigating FBR. In various implementations, materialsfor constructing the membrane layer 277 include gels comprising proteinssuch as albumin and collagen, as well as non-proteinaceous polymers suchas polyacrylamide, poly(ethylene oxide) poly(hydroxyethylmethacrylate)and its derivatives, and other hydrophilic polymers and copolymers, inboth crosslinked and non-crosslinked form.

The spout aperture diameter 272 in part controls the effectiveoperational membrane permeability ratio. In the exemplary embodiment,the aperture diameter is correlated to the range of concentration thetarget analyte (e.g., glucose) that can be detected by the detectorelement. A larger aperture diameter corresponds to a lower permeabilityratio of oxygen to glucose, and hence a greater sensitivity to a givenconcentration of glucose within the tissue proximate the aperture (andtherefore a lower minimum concentration that can be accuratelydetected). However, with the larger aperture, the detector will“saturate” more rapidly at a given oxygen concentration, and hence theupper bound of detection is similarly reduced. Conversely, a smallerdiameter aperture corresponds to an increased permeability ratio, andhence a higher minimum effective sensitivity (and corresponding highermaximum detectable concentration before saturation is reached).

It is also appreciated that the (i) in various embodiments, the aperture276 of the spout region 270 may be virtually any geometric shape so longas the desired permeability ratio is achieved, such as, for example,round, oval, elliptical, rectangular, triangular, star shaped, square,polygonal, and the like (see FIG. 2B), or even irregular, although round(circular) apertures are generally preferred because such shapes aremore amenable to manufacturing; and (ii) diameter is straightforwardlyrelated to area, and the present disclosure contemplates that area maybe a useful measure of “spout size” as it relates to adjusting theoperational characteristics of the detector element in place ofdiameter.

Moreover, other dimensional parameters have been identified by theinventors hereof as having an impact on detector element performance,and being a means by which such performance can be adjusted or optimizedas desired. For example, in addition to diameter of the aperture 276,the placement of the aperture(s) relative to the base of the cavity(e.g., height of the aperture above the base), the height 275 of thebase region above the underlying inner membrane (see FIG. 2), and eventhe vertical height of the sidewalls of the aperture itself 274 (seeFIG. 2A) can significantly affect the operation of the detector,including especially its response rate or detection time. See, e.g.,U.S. Pat. No. 7,336,984 and entitled “Membrane and Electrode Structurefor Implantable Sensor,” previously incorporated herein, which describesexemplary values (and techniques of determining such values) for animplantable glucose sensor of the type described herein.

Similarly, the diameter 273 of the base region (i.e., that regionunderlying the outer membrane 230; see FIG. 2) can affect operation ofthe detector, including specifically the range of concentrations ofanalyte that can be measured.

Hence, the present disclosure contemplates use of one or more of theforegoing dimensional parameters to configure or optimize the operationof the detector element 106 for a prescribed application.

As will be apparent to those skilled in the art, the outer(non-enzymatic) membrane layer 277 can be formed in any number ofdifferent ways. In the exemplary embodiment (see discussion of FIG. 3below), the non-enzymatic layer 277 is in effect “pour filled” into theaperture 276 of the outer housing membrane 230 atop the (crosslinked)enzymatic membrane matrix 240. However, the present disclosurecontemplates other techniques for formation, including for exampleprovision of a pre-formed membrane 277 which is inserted into theaperture in operative contact with the enzyme material 240, or evenchemical or other treatments of the upper surface of enzyme material240, including various de-immunizing treatments. In all cases, it isrequired that the outer membrane layer 277 be sufficiently permeable toanalytes and co-reactants to enable correct operation of the detector.In the exemplary embodiment, the outer membrane 277 comprises acrosslinked albumin, which exhibits the aforementioned desirableproperties of (i) lack of FBR-inducing enzymes, (ii) non-porosity, and(iii) electrical insulation. Notably, the exemplary albumin materialused for the membrane layer 277 is biocompatible; in the present context(a tissue-located implant), the term ‘biocompatible’ as applied to themembrane layer 277 indicates that the material itself does notsignificantly exacerbate the FBR which is otherwise expected to occurwith any implant. So, the amount/degree of fibrous tissue formation thatresults from the FBR (which nonetheless occurs due to natural bodyprocesses) is advantageously minimized, compared to what might beobtained with another less-biocompatible material.

Notably, in the exemplary implementation, the hydrophilic albumin of theouter membrane 277 is in direct contact with the (hydrophilic) tissue ofthe host, thereby advantageously providing a “like-to-like” interface,which also contributes to the stability of the detector elements overtime due to, among other things, the aforementioned non-exacerbation ofFBR or other host responses.

It is also noted that the exemplary membrane layer 277 described herein,by virtue of its non-exacerbation of FBR in the host (e.g., through useof a biocompatible material such as crosslinked albumin), furtherresults in mitigation of the formation of significant fibrous tissueresponse, which could otherwise interfere with optimal operation of thesensor detector elements or reduce their accuracy due to, inter alia,reduced blood vessel density in the fibrous tissue. So, in effect, thenon-exacerbation of FBR and non-encouragement of blood vessel ingrowthinto the membrane layer 277 by the exemplary embodiment herein actually(and somewhat counter-intuitively) stabilizes blood perfusion and bloodglucose delivery to the detector elements, and avoids having tosecond-guess the largely unpredictable modulation process, especiallyover longer periods of implantation.

Other biostable polymers suitable as outer (housing) membrane materialsinclude, for example, hydrophilic polyurethanes, silicones,poly(hydroxyethylmethacrylate)s, polyesters, polyalkyl oxides(polyethylene oxide), polyvinyl alcohols, and polyethylene glycols andpolyvinyl pyrrolidone. See, inter alia, U.S. Patent ApplicationPublication No. 2013/0197332 previously incorporated herein, for adiscussion of other various outer membrane materials.

It will also be appreciated that the “vertical” spout/aperture profilecan take on many forms as dictated by a given application. See e.g.,FIGS. 2C-2F, wherein several exemplary configurations (shown incross-section) of apertures and enzyme material 240 are shown. As willbe appreciated by those of ordinary skill given this disclosure, theconfiguration of the aperture, including its placement relative to othercomponents within the detector element (including outer membrane 230,inner membrane 220, etc.), can affect the operation of the detectorelement. For example, the present disclosure contemplates that a roundedor smoothed and progressively narrowing aperture/spout shape (see FIG.2C) as one embodiment.

Likewise, a tapered or chamfered aperture (either “taper up” or “taperdown,” or both; see FIGS. 2E and 2F) may be useful in certaincircumstances. Notably, the “taper up” embodiment of FIG. 2E provides anadded advantage relating to mechanical stability; i.e., the taper of thenon-enzymatic membrane 277 and sidewalls of the aperture 276 cooperateto retain the membrane 277 (once cured) in place, and resist dislocationor movement potentially resulting from e.g., thermal expansion of theunderlying enzyme matrix 240, or interaction between the outer surfaceof the membrane 277 and the host tissue/encapsulation FBR which mighttend to draw the membrane 277 out of the aperture 276. It will beappreciated that other shapes may similarly be used to provide suchmechanical stability, such as e.g., an inverted “T” shape (not shown).

As yet another example, a “rounded edge” or bull-nosed configuration ofthe aperture (FIG. 2D) may be useful.

Yet other configurations will be recognized by such skilled artisanswhen given the disclosure. These various vertical aperture/spoutprofiles may further be combined with one or more of the “horizontal”planar aperture shapes shown in FIG. 2B (or yet others) in order toachieve the desired performance attributes. It will also be appreciatedthat the thickness of the non-enzyme layer or membrane 277 may also bevaried (as can other properties of the non-enzymatic membrane 277, suchas density or permeability) so as to effect the desired rate ofpermeation of the analyte and any associated coreactant(s) (e.g.,glucose and oxygen) through the membrane, and also to frustrate orcontrol blood vessel growth into the membrane 277.

Yet further, it is recognized that the membrane 277 need not have aconsistent shape or thickness; e.g., it may have thicker or narrowerregions (which may or may not be symmetrical), such as to create regionsof e.g., greater or lesser permeation relative to the underlyingenzyme-containing matrix 240. In one such variant, the edges of themembrane 277 are made thinner than the central portion, such that moreanalyte/oxygen permeates through the outer regions per unit time, andhence diffuses to the outer regions of the matrix 240 (i.e., those whichare not directly under or proximate to the aperture 276). Hence, a more“even burn” of enzyme material is achieved.

Likewise, it will be appreciated that the “fill level” of enzymematerial 240 relative to the spout/aperture can be varied. For example,as shown in the embodiment of FIG. 2A, the top of such enzymaticmaterial 240 is roughly coincident with the height of the bottom of theaperture 276. The foregoing variation in fill level must, however, beconsistent with the requirement that at least a portion of the outer(non-enzyme) layer or membrane 277 generally must be in physical orchemical “contact” with the underlying enzyme material 240 (absent anyinterposed material or vehicle) so as to permit permeation of theanalyte(s) (e.g., glucose, and free oxygen) into the enzyme material240. It will be recognized that such “chemical contact” may take variousforms, including direct physical contact, indirect contact via aninterposed layer of material or fluid (such as to promote binding of theouter membrane 277 to the matrix 240) which does not substantiallymitigate permeation, or yet other ways.

In another variant, the outer membrane 277 may be directly bonded orattached to the underlying matrix (whether enzymatic, such as in theprimary detector elements, or non-enzymatic, such as in the secondarydetector elements). For example, in one implementation, crosslinkedalbumin is used in the outer membrane 277, and is bonded to theunderlying crosslinked albumin-containing enzymatic matrix 240, so as topromote inter alia, constant and complete glucose and oxygen moleculemigration during operation. Further, the direct bonding of the two citedmembrane layers helps to ensure a stable mechanical structure of themembrane assembly, a prerequisite in ensuring stable, predictable sensorresponse characteristics. In one such implementation, the two layers(outer membrane 277 and matrix 240) are bonded via a chemicalcrosslinking agent such as glutaraldehyde, which, for example, when thelayers comprise proteinaceous materials, promotes chemical bondingbetween the layers as well as crosslinking within each layer.

Moreover, as previously indicated, the outer membrane layer 277 may alsobe (chemically) bonded to the outer silicone membrane shell 230 such asat the interface between the outer edge of the membrane layer 277 andthe inner periphery of the aperture 276. This approach can help furtherprotect against any migration of the enzymes in the matrix 240 outwardtoward the host tissue, thereby avoiding any exposure thereof (andpossible further FBR to the enzyme(s)).

Additionally, it will be appreciated that while in various embodiments,the exemplary spout region 270 is filled or layered with an additionalnon-enzyme material or membrane 277 (such as to reduce the immunogenicpotential of the enzymatic material 240), such layer or membrane 277 maynot be required in certain cases. For instance, where a given detectordoes not utilize any enzymatic material (or uses an enzymatic materialthat produces limited if any FBR in the host tissue, it may be feasibleto eliminate the outer non-enzymatic layer). Moreover, where the spoutaperture(s) 276 has a comparatively small diameter, such as where aplurality of small apertures 276 are used in place of a single largeraperture, direct contact surface area with host tissue may be quitesmall and spatially distributed, thereby potentially obviating the needfor the “buffer” membrane 277.

It is also envisioned that a spatial gradient in enzyme concentrationwithin the enzyme material can be employed, such that e.g., theconcentration is reduced proximate the host solid tissue, therebyostensibly mitigating FBR due to irritation by the enzymes, transientperoxides, etc. Further, it is contemplated that a chemical treatmentapplied during manufacturing could be employed to “de-immunize” anexposed surface of membrane material 240, obviating the need for thebuffer membrane 277.

Heterogeneous Detector Element Arrays

It is contemplated that in other embodiments, the detector array 104includes detector elements 106 with different spout (aperture) diametersand/or other physical characteristics; e.g., one detector or set ofdetectors with larger spout diameters and/or heights, and anotherdetector/set with smaller spout diameters and/or heights. Havingmultiple detector elements 106 with such different physicalcharacteristics (and hence operating characteristics) is beneficial forany number of reasons, including maintaining a broader desired sensorresponse range.

For example, where the variation of the concentration of the underlyinganalyte being measured is substantial (whether spatially or over time),there is the possibility that a detector or set of detectors with e.g.,a common, finite range of detection may be “over-ranged” or“under-ranged” such that it/they are incapable of accurately detectingthe concentration through such a broad range of levels.

In one implementation (see FIG. 3), twelve (12) sensor elements areincluded on the sensor apparatus 300, two (2) of which 306 a areconfigured to measure glucose level within a first response range, twoof which 306 c are configured to measure glucose level within a second(at least partly differing) response range, and two of which 306 e areconfigured to measure glucose level within a third range which is atleast partly different from the first and second ranges. The remainingsix (6) sensor elements 306 b, 306 d, 306 f are used to measure oxygenconcentration (i.e., are reference elements).

Moreover, it will be appreciated that most any sensor will tend to havea detection “sweet spot,” wherein the operation of the sensor(s) (e.g.,its signal-to-noise ratio and therefore its accuracy) are optimized ascompared to operation at other values, such as those at the ends of itsdynamic range. Hence, it may be desirable to use that particulardetector for measuring analyte concentrations that fall at or near thesweet spot, so as to provide the most accurate results. Having two ormore heterogeneous detector elements with differing or staggered sweetspots (such as the apparatus 300 of FIG. 3) thereby enables moreaccurate measurement over a broader range than if a single detector(range) was used.

Accordingly, another exemplary embodiment of the sensor apparatusdescribed herein may include either or both of: (i) multiple detectorelements with respective “staggered” ranges/rates of detection operatingin parallel (as in the apparatus of FIG. 3), and/or (ii) multipledetector elements with respective “staggered” ranges/rates of detectionthat are selectively switched on/off in response to, e.g., the analyteconcentration reaching a prescribed upper or lower threshold.

In one such embodiment, the sensor apparatus 300 includes two or moresets of detector elements 306 having different ranges of detection, andassociated control logic such that the output of the various detectorelements can be selectively utilized while the apparatus 300 isimplanted in vivo. In one such approach, each detector element includesa pre-designated upper and lower threshold value for analyteconcentration sensitivity, such that operation of the particulardetector element outside of those bounds is less desirable (or eveninoperable). In one implementation, the detector physical attributes(e.g., aperture diameter, base height, etc.) described above for eachsuccessive detector are coordinated such that the upper and lower boundsof each are generally contiguous, thereby forming a “stitched together”virtual sensor with expanded range of detection. The supportingcircuitry of the sensor apparatus (or alternatively, off-sensor logicsuch as on a user's wireless monitor) is configured in one variant to:(i) determine a trend or slope of analyte concentration, and (ii) aproximity to a given threshold for a given detector element (or set ofelements), such that the circuitry can “hand off” from one detectorelement/set (e.g., with a first operational range or sensitivity) to asecond element/set with a contiguous or overlapping range orsensitivity), thereby extending the dynamic range of the device as awhole.

Moreover, the aforementioned upper/lower thresholds or bounds can beselected such that the aforementioned “sweet spot” of the particulardetector element is primarily used, with handoff to another element/setoccurring before significant degradation of performance occurs. Hence,in one scenario, the upper and lower thresholds of a first sensorapparatus 300 with say six (6) heterogeneous, staggered sets of workingand reference detectors can be adjusted to “stitch together” the samedynamic range of a similar homogenous detector sensor apparatus (i.e.,four sets of identical detector elements, such as the apparatus 100 ofFIG. 1), yet with increased accuracy throughout the range, since each ofthe constituent detector elements in the heterogeneous apparatus 300 areeach operating only in their “sweet spot” before handing off to the nextdetector element.

The present disclosure further contemplates that such thresholds orbounds: (i) can be selected independent of one another; and/or (ii) canbe set dynamically while the apparatus 300 is implanted. For example, inone scenario, operational detector elements are continuously orperiodically monitored to confirm accuracy, and/or detect anydegradation of performance (e.g., due to equipment degradation,progressive FBR affecting that detector element, etc.); when suchdegradation is detected, affecting say a lower limit of analyteconcentration that can be detected, that particular detector element canhave its lower threshold adjusted upward, such that handoff to anotherelement capable of more accurately monitoring concentrations in thatrange.

Alternatively, each of the aforementioned heterogeneous sensor sets 306a-f may simply be operated in parallel, and data generated by eachtransmitted off-device (e.g., via wireless interface to an externalreceiver) for subsequent processing of the raw data on the externalreceiver device or on an external computational platform, such as viaapplication software running on a personal computer or server andconfigured to identify the most optimal data from each sensor set 306within the “raw” data generated by that sensor set and transmittedoff-device, and utilize the identified optimal data to provide arepresentation of the measured analyte concentration over the entirerange of values encountered, ostensibly with greater accuracy than thatprovided by a comparable homogenous detector configuration. Forinstance, in one such implementation, the sensor sets 306 are evaluatedat e.g., time of manufacture (or statistically modeled) so that the“sweet spot” of each particular set on a given device 300 is known apriori; such evaluation or modeling data is utilized by theaforementioned application software to filter data obtained from the insitu device 300 so as to retain only data associated with measuredglucose concentrations falling within the optimal range of eachparticular sensor set 306.

Methods for Manufacturing

In another aspect, methods of manufacturing one or more sensor apparatus100, 300 of the present disclosure are described in detail. Referringnow to FIG. 4, one exemplary embodiment of a method 400 of manufacturingthe various membranes and/or layers of the exemplary detector elementincluding the novel spout region is disclosed.

In the illustrated embodiment, the method 400 includes first providing asubstrate (e.g. ceramic or similar) per step 402. Next, the working,reference and counter electrodes (as applicable) 217, 218, 219 andassociated feedthroughs are formed within the substrate (in oneembodiment as previously described herein with reference to U.S. Pat.No. 8,763,245 to Lucisano et al. entitled “Hermetic feedthrough assemblyfor ceramic body”) per step 304.

Next, per step 406, an electrolyte layer is formed over at least aportion of the substrate, including at least a portion of theelectrodes.

An inner (e.g., polymer, oxygen-permeable) membrane 220 is next formedover the electrolyte layer per step 408. In one embodiment, the innermembrane comprises a silicone rubber compound, although it will beappreciated that other materials may be used consistent with the presentdisclosure.

Per step 410, the outer hydrophobic membrane 230 is next formed(although this can also be formed concurrently with the inner membrane),such as via molding of a silicone rubber compound identical or similarto that used for the inner membrane in the desired shape and dimensions,including the aperture 276. Advantageously, the mold(s) used for formingthe outer membrane 230 can easily be modified or adjusted (or multiplemolds used), such that detectors with different operatingcharacteristics can readily be produced (including variants where allother components remain the same).

The outer membrane 230 is formed or disposed over the inner membrane perstep 412 such that the inner and outer membranes form a cavity toencapsulate the enzyme material 240. In the exemplary embodiment, theinner membrane 220 and outer membrane 230 are joined together via anadhesive (e.g., room temperature vulcanizing (RTV) rubber adhesive) orother bonding process, although it is also appreciated that the outerhydrophobic membrane and the inner membrane may potentially be formed asa common component (i.e., one piece) when the materials selected foreach are the same.

Next, per step 414, the enzyme matrix material 240 is formulated andinserted into the cavity 271 such that it at least contacts the innermembrane and aperture (i.e., to the desired level), and then cured(e.g., via the introduction of added chemical cross-linking agent) toeffectuate the desired degree of crosslinking (and enzymeimmobilization). In the exemplary embodiment, the enzyme matrix material240 includes the enzyme components (i.e., catalase, oxidase), as well asa binder protein (albumin), all dissolved in an aqueous buffer(phosphate-buffered saline), and also a small percentage by volume of achemical cross-linking agent such as glutaraldehyde. The resultingmixture is in a substantially liquid or flowable form beforeintroduction into the cavity formed between the inner and outermembranes 220, 230. Advantageously, use of the buffer and thecross-linking agent within the liquid/flowable enzyme material mixturehelps reduce or eliminate formation of voids or “bubbles” within thematerial after curing (discussed below), thereby enhancing theperformance of the sensor element after implantation.

Moreover, in certain embodiments, it is desired to maintain at leastportions of the side surfaces of the aperture 276 substantiallyenzyme-free, so as to inter alia, mitigate the chances of any enzymematerial coming in contact with the host's surrounding tissue (i.e.,after implantation), such contact potentially resulting in undesired FBRdue to exposure to the enzymes. Hence, in one implementation, the enzymematerial mixture 240 is used to fill the cavity 271 up to a levelcoincident with the bottom edge of the side walls of the aperture 276,and the enzyme material is prevented from contacting the sidewallsduring such fill.

Next, per step 416, the enzyme material 240 within the cavity 271 is“cured,” such as via the application of additional chemicalcross-linking agent atop the material 240 via the aperture 276, or bydiffusion through the outer membrane 230. It will be appreciated,however, that while chemical cross-linking is described herein withrespect to the exemplary embodiments, the disclosure contemplates othermeans of curing the matrix material, including e.g., via heat and/orradiation, whether alone or in combination with the aforementionedchemical agents.

After the curing (e.g., crosslinking) of the enzyme matrix material iscompleted, the non-enzyme membrane material is formulated and introducedinto the aperture region 276 (step 418). In the exemplary embodiment,the non-enzyme membrane material is also in a substantially liquefied orflowable form and includes a protein such as an albumin (e.g.,recombinant human albumin), and a portion of the aforementionedbuffering agent (although different/heterogeneous buffering agents maybe used in the enzymatic and non-enzymatic membranes if desired).Addition of the crosslinking agent prior to the filling procedure hasnot been found advantageous with the albumin material, therefore itsinclusion prior to filling is not required.

Once the non-enzyme membrane material is disposed within the aperture tothe desired height (e.g., approximately even with the top surface of theouter membrane 230 proximate the aperture 276), the non-enzyme materialis cured (e.g., via chemical cross-linking similar to that used for theenzymatic material, or other processes) per step 420. This processfurther causes bonding between the bottom portion of the non-enzymaticmembrane later and the top of the (previously cured) enzymatic material240 at least in the region of the aperture 276. Advantageously, suchbonding helps both avoid the formation of gaps or voids between thelayers, and ensures consistent oxygen and glucose migration from thenon-enzymatic membrane to the enzyme material during operation.

It will be recognized that while certain embodiments of the presentdisclosure are described in terms of a specific sequence of steps of amethod, these descriptions are only illustrative of the broader methodsdescribed herein, and may be modified as required by the particularapplication. Certain steps may be rendered unnecessary or optional undercertain circumstances. Additionally, certain steps or functionality maybe added to the disclosed embodiments, or the order of performance oftwo or more steps permuted. All such variations are considered to beencompassed within the disclosure and claimed herein.

While the above detailed description has shown, described, and pointedout novel features as applied to various embodiments, it will beunderstood that various omissions, substitutions, and changes in theform and details of the device or process illustrated may be made bythose skilled in the art without departing from principles describedherein. The foregoing description is of the best mode presentlycontemplated. This description is in no way meant to be limiting, butrather should be taken as illustrative of the general principlesdescribed herein. The scope of the disclosure should be determined withreference to the claims.

What is claimed is: 1.-31. (canceled)
 32. A method of manufacturing ananalyte detector element, comprising: embedding at least one electrodewithin a substrate; disposing an electrolyte over at least a portion ofthe substrate, the at least portion of the substrate comprising at leasta portion of the electrode; forming a membrane structure over theelectrolyte, the membrane structure comprising a cavity and an aperturecommunicating with the cavity; disposing an enzyme matrix material intothe cavity via the aperture such that the enzyme matric materialcontacts at least a portion of an interior wall of the membranestructure; and disposing a non-enzymatic material at least partly withinthe aperture such that the non-enzymatic material contacts at least aportion of the enzyme matrix material.
 33. The method of claim 32,wherein the disposing of the enzyme matrix material into the cavitycomprises: disposing a flowable enzyme matrix material within thecavity; and subsequently cross-linking at least a portion of theflowable matrix material while disposed within the cavity.
 34. Themethod of claim 33, wherein the flowable enzyme matrix material at leastin part comprises a chemically cross-linkable substance; and thesubsequent cross-linking comprises applying, via the aperture, aflowable cross-linking agent onto the flowable enzyme matrix materialdisposed in the cavity, thereby forming a crosslinked enzymatic matrixmaterial.
 35. The method of claim 34, wherein the disposing of thenon-enzymatic material comprises disposing, via the aperture, a flowablenon-enzyme matrix material atop the crosslinked enzymatic matrixmaterial.
 36. The method of claim 35, wherein the disposing of thenon-enzymatic material further comprises chemically cross-linking thenon-enzymatic material within the aperture via applying a cross-linkingagent to at least a top surface of the non-enzymatic material disposedwithin the aperture.
 37. The method of claim 36, wherein the chemicalcross-linking of the non-enzymatic material within the aperture at leastin part comprises forming a porous outer membrane within the aperture,the porous outer membrane having a porosity greater than that of themembrane structure.
 38. The method of claim 36, wherein the chemicalcross-linking of the non-enzymatic material within the aperture at leastin part comprises causing formation of a bond between a top surface ofthe crosslinked enzymatic matrix and a bottom surface of thenon-enzymatic material disposed within the aperture.
 39. The method ofclaim 36, wherein the chemical cross-linking of the non-enzymaticmaterial within the aperture at least in part comprises causingformation of a bond between one or more wall surfaces of the membranestructure defining the aperture and the non-enzymatic material disposedwithin the aperture.
 40. The method of claim 32, wherein the forming ofthe membrane structure over the electrolyte comprises: forming an innersilicone rubber-based membrane over at least a portion of theelectrolyte; forming an outer silicone rubber-based membrane having theaperture formed therein; and adhering the outer silicone rubber-basedmembrane to the inner silicone rubber-based membrane such that the innerand outer silicone rubber-based membranes define the cavity.
 41. Themethod of claim 32, wherein the disposing the flowable enzyme matrixmaterial into the cavity comprises disposing so as to avoidcontamination of one or more wall surfaces of the membrane structuredefining the aperture by the flowable enzyme matrix material.
 42. Themethod of claim 41, wherein the disposing so as to avoid contaminationcomprises filling the cavity to a level coincident with a bottominterior edge of the one or more wall surfaces of the membrane structuredefining the aperture, the filling comprising filling so that none ofthe flowable enzyme matrix material contacts the one or more wallsurfaces.
 43. A method of manufacturing an analyte detector element,comprising: disposing an electrolyte material over at least a portion ofan active surface of an electrode; forming a hydrophobic membranestructure over the electrolyte material, the hydrophobic membranestructure comprising at least (i) a cavity and (ii) a spout region incommunication with the cavity; forming an enzyme matrix within thecavity such that the enzyme matrix contacts at least a portion of abottom interior wall of the cavity; and forming a hydrophilic membraneat least partly within the spout region such that the hydrophilicmembrane contacts at least a portion of a top surface of the enzymematrix.
 44. The method of claim 43, wherein the forming of the spoutregion at least in part comprises forming a specified three-dimensionalshape of the spout region, the specified three-dimensional shapeconfigured to regulate analyte diffusion across the hydrophilicmembrane.
 45. The method of claim 43, wherein the forming of the enzymematrix at least in part comprises: disposing a flowable enzyme matrixmaterial within the cavity, the flowable enzyme matrix material at leastin part comprising a first chemically cross-linkable material; applyinga first chemical crosslinking agent to the flowable enzyme matrixmaterial within the cavity, thereby causing: cross-linking of the firstchemically cross-linkable material, and forming of a cross-linked enzymematrix disposed within the cavity and at least partly coincident with abottom edge of an inner wall of the spout region.
 46. The method ofclaim 45, the forming of the hydrophilic membrane at least in partcomprises: subsequent to the applying of the first chemical crosslinkingagent and the forming of the cross-linked enzyme matrix, disposing aflowable hydrophilic membrane material at least partially within thespout region, the flowable hydrophilic membrane material at least inpart comprising a second chemically cross-linkable material; andapplying a second chemical cross-linking agent, the second chemicalcrosslinking agent causing: cross-linking of the second chemicallycross-linkable material; forming of a cross-linked hydrophilic membranedisposed within the spout region and substantially coincident with a topedge of an outer wall of the spout region; and bonding of a bottomsurface of the cross-linked hydrophilic membrane to a top surface of thecross-linked enzyme matrix.
 47. The method of claim 46, wherein thedisposing a flowable enzyme matrix material within the cavity comprises:filling the cavity with the flowable enzyme matrix material through anaperture of the spout region; and preventing contact of the flowableenzyme matrix material with each of the outer wall of the spout regionand one or more interior side walls of the aperture.
 48. The method ofclaim 46, wherein the forming of the cross-linked hydrophilic membranedisposed within the spout region comprises sealing of the cross-linkedenzyme matrix within the cavity.
 49. A method of manufacturing ananalyte detector, comprising: seating one or more electrodes within adetector substrate; disposing an electrolyte material over at least aportion of an active surface of each of the one or more electrodes;forming a membrane structure over the electrolyte material, thehydrophobic membrane structure comprising at least a cavity and anaperture communicating with the cavity, the cavity disposed over atleast one of the one or more electrodes; disposing a flowable enzymematrix material within the cavity via the aperture while substantiallypreventing contamination of an outer surface of the membrane structureand one or more inner walls of the aperture, the flowable enzyme matrixmaterial at least in part comprising a first cross-linkable material andone or more classes of enzymes; applying a first crosslinking agent tothe flowable enzyme matrix material disposed in the cavity, therebyforming a cross-linked enzyme matrix; and disposing a non-enzymaticmaterial within the aperture; and forming a seal between thenon-enzymatic material and the at least one inner wall of the aperture,the seal configured to substantially cause enclosure of the cross-linkedenzyme matrix within the cavity.
 50. The method of claim 49, wherein thedisposing of the non-enzymatic material within the aperture at least inpart comprises disposing a flowable non-enzymatic material within theaperture, the flowable non-enzymatic material at least in partcomprising a second cross-linkable material; and the forming of the sealbetween the between the non-enzymatic material and the at least oneinner wall of the aperture at least in part comprises applying a secondcross-linking agent to the flowable non-enzymatic material within theaperture thereby forming a cross-linked non-enzyme material bonded tothe at least one inner wall of the aperture.
 51. The method of claim 49,wherein the disposing of the flowable enzyme matrix material within thecavity at least in part comprises filling the cavity with the flowableenzyme matrix material such that a top surface of the enzyme matrixmaterial is substantially coincident with an internal edge of the atleast one inner wall of the aperture; and the disposing of thenon-enzymatic material within the aperture at least in part comprisesfilling the aperture with the non-enzymatic material such that a topsurface of the non-enzymatic material is within a prescribed distance ofan external edge of the at least one inner wall of the aperture.